Atlas of Assisted Reproductive Technologies

Preface
Acknowledgments
Contents
Part I: ART Laboratory and Equipment
Set Up of an IVF Lab
1 Non-Sterile Area
2 Sterile Area
3 Infrastructure of the IVF Laboratory
4 HEPA Filters (High-Efficiency Particulate Air) (Fig. 22)
5 Air Handling Unit (AHU) (Fig. 23)
References
Equipment for Micromanipulation
1 ICSI Instrumentation
1.1 Main Components
1.2 Types of Micromanipulators: Micromanipulators can be of three types
1.3 Function of Manipulators
1.3.1 Hydraulic Micromanipulator Components and Their Function
1.4 Eppendrof Micromanipulator: Fully Motorized
References
Part II: Clinical Procedures
Ultrasound Imaging of the Ovary in Infertility
References
Oocyte Retrieval
1 Equipment and Consumables Required for Ovum Pick Up
2 Patient Preparation and Anaesthesia
3 Preparation of the Ultrasound Machine, the Probe and the Oocyte Retrieval Set
4 Transvaginal Oocyte Retrieval Technique
5 Post Procedure Care
6 Complications
7 Other Routes for Oocyte Retrieval
8 Dos and Dont’s
9 Debatable Issues
10 Quality Assurance and Performance
11 Conclusion
References
Embryo Transfer
1 Introduction
1.1 Embryo Transfer Techniques
2 Scheduling and Consents
2.1 Day of Transfer
2.2 No. of Embryos to be Transferred
2.3 Recommendations for the Limit to the Number of Embryos to Transfer
2.4 Grading of Embryos
2.5 Blastocyst Grading Istanbul Consensus
3 Trial Transfer and Difficulty Assessment
4 Embryo Transfer Procedure
4.1 Setup for ET OT or ET Suit
4.2 Setup for ET Embryology Laboratory
4.3 Equipment and Disposable
4.4 ET Catheter
4.5 ET Transfer Procedure
4.5.1 IVF Lab Procedure
4.6 Loading Techniques
4.7 Embryo Transfer Clinical Procedure
4.8 Remaining Embryos
References
Intrauterine Insemination
1 Follicle Monitoring and Timing of IUI
2 Sample Collection
3 Semen Analysis
4 Semen Processing
5 Procedure
6 Post Procedure Instructions
7 Types of IUI Catheters
8 Strategies to Overcome Difficulties in IUI
9 Double Vs. Single IUI
References
Sperm Retrieval Techniques
1 Introduction
2 Sperm Retrieval Techniques
2.1 Materials, Equipment, and Reagents
2.1.1 Operating Room
2.1.2 IVF Laboratory
2.1.3 Laboratory Setup
2.2 PESA
2.3 MESA
2.4 TESA
2.5 Testicular Sperm Extraction (TESE)
2.6 Micro-TESE
3 Surgical Complications
4 Results
References
Glossary
Part III: Gametes: Sperm and Oocyte
Oocyte Morphology
1 Granularity in Cytoplasm
2 SER
3 Cytoplasmic Inclusion
4 Vacoules
5 Shape
6 Zona Pellucida Abnormalities
7 Perivitteline Space Abnormalities
8 Polar Body
References
Semen Analysis
References
Human Sperm Morphology
1 Slide Preparation
2 Normal Spermatozoa
2.1 Normal Spermatozoa
2.2 Abnormal Spermatozoa
3 Strict Criteria
3.1 Head Defects
3.1.1 Large Head
3.1.2 Microcephaly/Small Sperm Head
3.1.3 Amorphous Head
3.1.4 Tapered Head
3.1.5 Pyriform Head: (Fig. 7)
3.1.6 Small Acrosomal Area
3.2 Neck/Mid-Piece Defect
3.2.1 Bent Mid-Piece
3.2.2 Over- Thick Mid-Piece
3.2.3 Cytoplasmic Droplet
3.3 Tail Defect
3.3.1 Coiled Tail
3.3.2 Tail-Tip Coiling
3.3.3 Double Tail
References
Assessment of Sperm DNA Damage
Retrograde Ejaculation
1 Part 1. Introduction
2 Part 2. Signs of Retrograde and Clinical Investigation
3 Part 3. Laboratory Confirmation of Retrograde Ejaculation
3.1 Patient Preparation
3.2 Laboratory Diagnosis (Fig. 3)
4 Part 4. Preparation of Retrograde Samples for ART
References
Part IV: Embryo
Embryo Morphology and Embryoscopy
1 Cleavage Stage Embryo
2 Compaction
3 Blastocyst
4 Morphology Vs. Morphokinetics
References
The Blastocyst
1 Expansion Grades of a Blastocyst
2 Inner Cell Mass and Trophectoderm
3 Blastocyst with Excluded Cells or “Fragmentation”
4 Abnormally Shaped Blastocyst
5 Site of Hatching of Blastocyst
6 Expansion and Contraction of a Blastocyst
7 Cytoplasmic Strings in Blastocyst
References
Part V: Embryology Procedures
Semen Preparation Techniques
1 Introduction
2 Collection of Semen
3 Semen Analysis Before Preparation
4 Sperm Preparation Methods
4.1 Swim-Up
4.1.1 Layering and Swim-Up
Media Used
Procedure
4.2 Density Gradient Centrifugation
4.2.1 Procedure
4.3 Magnetic Activated Cell Sorting
4.4 Microfluidic Sperm Sorting (MFSS)
4.4.1 Procedure
5 Conclusion
References
Cryopreservation
1 Sperm Cryopreservation
1.1 Principle
2 Semen Collection and Preparation of Sperm for Cryopreservation (Fig. 1)
3 Cryopreservation Technique
3.1 Slow Freezing
3.2 Rapid Freezing [4]
3.3 Thawing (Fig. 5)
4 Oocyte and Embryo Cryopreservation
4.1 Introduction
4.2 Principle
4.3 Steps of Vitrification
4.3.1 Virtification Steps
4.3.2 Oocyte in Equilibrium Medium
4.3.3 Oocyte in Vitrification Medium
4.3.4 Oocyte Loading
4.3.5 Four Cell Stage in Equilibrium Medium
4.3.6 Blastocyst Stage in Equilibrium Medium
4.3.7 Blastocyst Loading
4.3.8 Liquid Nitrogen Storage Tank
References
ICSI and Micromanipulation
1 Introduction
2 Equipment
3 Setting Up of a Micromanipulator System
4 Preparation of Gametes for ICSI
4.1 Sperm Preparation
4.2 Oocyte Preparation
5 Micromanipulation Procedures
5.1 ICSI
5.2 Assisted Hatching
5.3 Embryo Biopsy
6 Conclusion
References
Preimplantation Genetic Testing (PGT)
1 Introduction
2 Embryo Biopsy
2.1 PGT-A
2.2 PGT-SR
2.3 PGT-M
3 Future Perspectives
References
Preimplantation Genetic Testing for Aneuploidy (PGT-A): Lab Aspects
1 State-of-Art Lab Facility for Embryo Biopsy
2 LASER: Laser Is an Important Tool Required to Enable an Embryo Biopsy
2.1 New Lasers
2.2 Dynamic Laser
2.3 Static Laser
3 Micromanipulator
4 Diagrammatic Representation of Events in a PGT-A
5 Assisted Hatching for Embryo Biopsy
5.1 Chemical Hatching
5.2 Mechanical Hatching
5.3 Laser-Assisted Hatching
6 Various Stages of Biopsies
6.1 Polar Body Biopsy
6.2 Cleavage Stage Biopsy
6.2.1 Advantages of Cleavage Stage Biopsy
6.2.2 Limitations of Cleavage Stage Biopsy
6.3 Morula Stage Biopsy
6.4 Blastocyst Biopsy
6.4.1 Advantages of TE Biopsy
6.4.2 Limitations of TE Biopsy
7 Timing for Laser-Assisted Hatching (LAH) for Blastocyst Biopsy: Day-3 Vs. Day-5
8 Mosaicism
8.1 Recommendations for the Laboratory (if Reporting Mosaic Aneuploidies)
8.2 Recommendations for the Clinician
9 Grade of Embryo and Correlation with Aneuploidy
10 When to Freeze After Biopsy?
10.1 Cryopreservation of Embryos
References